title: High-resolution imaging of cytoplasmic Ca2+ dynamics in Arabidopsis roots.
abstract: This protocol describes a method for imaging cytoplasmic Ca(2+) dynamics in roots with high resolution using confocal laser scanning microscopy (CLSM). The Förster (fluorescence) resonance energy transfer (FRET)-based genetically modified Ca(2+) indicator Yellow Cameleon YC3.6, stably expressed in plants under the control of the ubiquitin promoter UBQ10, is used for Ca(2+) measurements. This protocol enables imaging of 5- to 7-d-old seedlings with high-magnification objectives (25×, 40×, and 63×).
authors.affiliation: Institute of Plant Biology and Biotechnology, University of Münster, 48149 Münster, Germany.
authors.forename: Smrutisanjita
authors.initials: S
authors.lastname: Behera
authors.forename: Jörg
authors.initials: J
authors.lastname: Kudla
mesh_terms: D017360:Arabidopsis / Q000737:chemistry*; D002118:Calcium / Q000032:analysis*; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002413:Cations, Divalent / Q000032:analysis; D003593:Cytoplasm / Q000737:chemistry*; D015870:Gene Expression; D017930:Genes, Reporter; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods*; D018517:Plant Roots / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D002135:Calcium-Binding Proteins; D002413:Cations, Divalent; C495639:yellow cameleon; D002118:Calcium